Protein Purification

Protein purification is a crucial aspect of all research projects in the Plant-Made Antibodies and Immunogens group.

Starting from a complex protein mixture (such as bacterial and yeast cultures, and plant leaf and seed extracts), proteins of interest are isolated in a suitable buffer and with a desired purity and concentration. All purifications are performed at 4 degrees C to preserve protein integrity.

Typical modes of protein purification include ion exchange, hydrophobic interaction, reversed-phase, size-exclusion and affinity chromatography. Often, several techniques are performed consecutively to obtain the final product. For example, an initial crude enrichment step by affinity chromatography can be followed by ion exchange chromatography for further protein enrichment or by a size-exclusion polishing step. The process is highly dependant on the specific protein characteristics,and hence requires customization.

Protein purification equipment: ÄKTAexplorer and ÄKTApurifier system (GE Healthcare).

Figure in project 4

Recommended Reading

De Buck, S., Nolf, J., De Meyer, T., Virdi, V., De Wilde, K., Van Lerberge, E., Van Droogenbroeck, B. and Depicker, A. (2013) Fusion of an Fc chain to a VHH boosts the accumulation levels in Arabidopsis seeds. Plant Biotechnol. J. 11, 1006-1016.

Virdi, V., Coddens, A., De Buck, S., Millet, S., Goddeeris, B.M., Cox, E., De Greve, H. and Depicker, A. (2013) Orally fed seeds producing designer IgAs protect weaned piglets against enterotoxigenic Escherichia coli infection. Proc. Natl. Acad. Sci. USA 110, 11809-11814.

Heijde, M., Binkert, M., Yin, R.H., Ares-Orpel, F., Rizzini, L., Van De Slijke, E., Persiau, G., Nolf, J., Gevaert, K., De Jaeger, G. and Ulm, R. (2013) Constitutively active UVR8 photoreceptor variant in Arabidopsis. Proc. Natl. Acad. Sci. USA 110, 20326-20331.